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Technical Data Sheet

PM1101

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PKC (α,β2,γ) M110
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms, Ck
82 kDa

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Western blot analysis of PKC isoforms in neonatal rat brain lysate. The rat brain blot was probed with anti-PKC (α,β,γ) at decreasing dilutions:

Lane 1 = 1:250
Lane 2 = 1:500
Lane 3 = 1:1000

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Immunocytochemical labeling of PKC relative to F-actin in chick DRG neurons. The cells were labeled with mouse monoclonal PKC (α,β,γ) antibody (PM1101), then detected using appropriate secondary antibody (Red). This labeling is compared to F-actin staining (Green). (Image provided by Dr. Gianluca Gallo at Drexel University).

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Application
Dilution


ELISA
1:2000


ICC
1:100


IP
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
The Protein Kinase C (PKC) family of homologous serine/threonine protein kinases is involved in a number of processes such as growth, differentiation, and cytokine secretion. At least eleven isozymes have been described. PKC consists of a single polypeptide chain containing four conserved regions (C) and five variable regions (V). The N-terminal half interacts with PKC activators Ca2+, phospholipid, diacylglycerol, or phorbol ester, while the C-terminal half contains the catalytic domain. The conventional PKC subfamily (α, β1, βII, and γ) is regulated by both Ca2+ and diacylglycerol. The PKC pathway represents a major signal transduction system that is activated following ligand-stimulation of transmembrane receptors by hormones, neurotransmitters and growth factors. The phosphorylation of multiple sites in conventional PKCs regulates their activity. In mast cells, FceRI stimulation leads to phosphorylation of tyrosine 658 and 662 of PKCα and PKCβI respectively. This phosphorylation requires autophosphorylation of serine 657 and 661 in these respective kinases.


Background References
Nishizuka, Y. (1988) Nature 334:661.
Kawakami et al. (2003) PNAS. USA 100:9470-9475.
Immunogen
Clone (M110) was generated from a recombinant human PKCγ that included amino acids residues 499-697.
Buffer and Storage
Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody detects 80-82kDa* proteins corresponding to the molecular mass of PKCα, PKCβ, and PKCγ on SDS-PAGE immunoblots of neonatal rat brain lysates. Similar results were observed in human and mouse lysates. Immunoprecipitation experiments with various PKC isoforms demonstrated this antibody detects PKCα, PKCβ2, and PKCγ, but does not detect PKCβ2 isoform. The antibody also detects these PKC isoforms in chick DRG neurons.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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